Fig3: Loss of Sirt3 decreases endothelial SOD2 activity and increases SOD2 expression without affecting other ROS-scavenging or generating systems. a Superoxide dismutase 2 (SOD2) activity based on superoxide dismutating capacity in HAEC following siRNA-mediated knockdown of Sirt3; enzymatic activity was normalized to SOD2 protein expression; medians and single data points are shown. b Overall SOD2 activity, as in a without normalizing to protein content. c SOD2 mRNA (left) and protein (right) expression in HAEC following siRNA-mediated knockdown of Sirt3 using quantitative PCR and western blot, respectively. d Acetylation of SOD2, precipitated from HAEC following transient knockdown of Sirt3 using western blot analysis. e Nitrosylation of SOD2, precipitated from HAEC following transient knockdown of Sirt3 using western blot analysis. f–h Expression analyses of f catalase, g SOD1, h SOD3, using quantitative PCR; beta actin served as loading control in western blots, representative blots are shown. At least three independent experiments in biological triplicates were performed, scr scrambled control

Fig3: Loss of Sirt3 decreases endothelial SOD2 activity and increases SOD2 expression without affecting other ROS-scavenging or generating systems. a Superoxide dismutase 2 (SOD2) activity based on superoxide dismutating capacity in HAEC following siRNA-mediated knockdown of Sirt3; enzymatic activity was normalized to SOD2 protein expression; medians and single data points are shown. b Overall SOD2 activity, as in a without normalizing to protein content. c SOD2 mRNA (left) and protein (right) expression in HAEC following siRNA-mediated knockdown of Sirt3 using quantitative PCR and western blot, respectively. d Acetylation of SOD2, precipitated from HAEC following transient knockdown of Sirt3 using western blot analysis. e Nitrosylation of SOD2, precipitated from HAEC following transient knockdown of Sirt3 using western blot analysis. f–h Expression analyses of f catalase, g SOD1, h SOD3, using quantitative PCR; beta actin served as loading control in western blots, representative blots are shown. At least three independent experiments in biological triplicates were performed, scr scrambled control